US 12,291,717 B2
RNA replicon for reprogramming somatic cells
Marco Alexander Poleganov, Mainz (DE); Mario Perkovic, Mainz (DE); Ugur Sahin, Mainz (DE); and Tim Beissert, Mainz (DE)
Assigned to BIONTECH RNA PHARMACEUTICALS GMBH, Mainz (DE); and TRON-Translationale Onkologie An Der Universitätsmedizin Der Johannes Gutenberg-Universität Mainz Gemeinnützige GMBH, Mainz (DE)
Appl. No. 16/645,707
Filed by BIONTECH RNA PHARMACEUTICALS GMBH, Mainz (DE); and TRON-TRANSLATIONALE ONKOLOGIE AN DER UNIVERSITÄTSMEDIZIN DER JOHANNES GUTENBERG-UNIVERSITAT MAINZ GEMEIN-NUTZIGE GMBH, Mainz (DE)
PCT Filed Sep. 11, 2018, PCT No. PCT/EP2018/074486
§ 371(c)(1), (2) Date Mar. 9, 2020,
PCT Pub. No. WO2019/053012, PCT Pub. Date Mar. 21, 2019.
Claims priority of application No. PCT/EP2017/073063 (WO), filed on Sep. 13, 2017.
Prior Publication US 2020/0277627 A1, Sep. 3, 2020
Int. Cl. C12N 15/86 (2006.01); C12N 5/074 (2010.01)
CPC C12N 15/86 (2013.01) [C12N 5/0696 (2013.01); C12N 2501/602 (2013.01); C12N 2501/603 (2013.01); C12N 2501/604 (2013.01); C12N 2501/605 (2013.01); C12N 2501/606 (2013.01); C12N 2501/608 (2013.01); C12N 2506/1307 (2013.01); C12N 2770/36143 (2013.01)] 19 Claims
 
1. A system comprising:
(i) an RNA construct for expressing a functional alphavirus non-structural protein, and
(ii) an RNA replicon comprising an open reading frame encoding a reprogramming factor and a 5′ replication recognition sequence, wherein the 5′ replication recognition sequence can be recognized by an alphaviral replicase in trans and is characterized in that it does not contain any initiation codons, wherein the 5′ replication recognition sequence does not overlap with a first open reading frame of the RNA replicon, and the initiation codon of the first open reading frame is in the 5′→3′ direction of the RNA replicon and is the first functional initiation codon; wherein
the RNA construct is an mRNA molecule comprising a 5′-cap; and
the RNA replicon does not comprise an open reading frame encoding a functional alphavirus non-structural protein.